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PMID: 7045809 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Laser crosslinking of E. coli RNA polymerase and T7 DNA.

Nucleic acids research ·Vol. 10 ·No. 7 ·1982-04-10 ·Pages 2399-414

Harrison CA, Turner DH, Hinkle DC

Abstract

The first photochemical crosslinking of a protein to a nucleic acid using laser excitation is reported. A single, 120 mJ, 20 ns pulse at 248 nm crosslinks about 10% of bound E. coli RNA polymerase to T7 DNA under the conditions studied. The crosslinking yield depends on mercaptoethanol concentration, and is a linear function of laser intensity. The protein subunits crosslinked to DNA are beta, beta' and sigma.

MeSH Terms
DNA, Viral/metabolism,radiation effects DNA-Directed RNA Polymerases/metabolism,radiation effects Escherichia coli/enzymology Kinetics Lasers Macromolecular Substances Protein Binding T-Phages/metabolism
Chemicals
DNA, Viral Macromolecular Substances DNA-Directed RNA Polymerases
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Harrison C A
Turner D H
Hinkle D C
References (22)
22 references, click to expand
  1. The mechanism of photochemical addition of cysteine to uracil and formation of dihydrouracil.
    Photochem Photobiol. 1970 May;11(5):349-59 PMID: 5429552
  2. Cleavage of structural proteins during the assembly of the head of bacteriophage T4.
    Nature. 1970 Aug 15;227(5259):680-5 PMID: 5432063
  3. Determination of molecular weight distributions of DNA by means of sedimentation in a sucrose gradient.
    Anal Biochem. 1969 Oct 15;32(1):14-30 PMID: 5398260
  4. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane.
    Biochemistry. 1971 Jun 22;10(13):2606-17 PMID: 4326772
  5. A mixed photoproduct of thymine and cysteine: 5-S-cysteine, 6-hydrothymine.
    Biochem Biophys Res Commun. 1970;39(6):1011-6 PMID: 5513241
  6. Studies of the binding of Escherichia coli RNA polymerase to DNA. I. The role of sigma subunit in site selection.
    J Mol Biol. 1972 Sep 28;70(2):157-85 PMID: 4562312
  7. Ultraviolet light-induced stable complexes of DNA and DNA polymerase.
    Biochim Biophys Acta. 1972 Nov 9;281(4):522-34 PMID: 4569287
  8. Induction of stable linkage between the deoxyribonucleic acid dependent ribonucleic acid polymerase and d(A-T)n-d(A-T)n by ultraviolet light.
    Biochemistry. 1974 Jan 29;13(3):485-93 PMID: 4589312
  9. Statistical time events in enzymes: a physical assessment.
    CRC Crit Rev Biochem. 1975 Aug;3(2):141-64 PMID: 1100316
  10. A procedure for the rapid, large-scall purification of Escherichia coli DNA-dependent RNA polymerase involving Polymin P precipitation and DNA-cellulose chromatography.
    Biochemistry. 1975 Oct 21;14(21):4634-8 PMID: 1101952
  11. Exposure of tryptophanyl residues in proteins. Quantitative determination by fluorescence quenching studies.
    Biochemistry. 1976 Feb 10;15(3):672-80 PMID: 1252418
  12. EcoRI endonuclease. Physical and catalytic properties of the homogenous enzyme.
    J Biol Chem. 1976 Oct 10;251(19):5866-74 PMID: 786985
  13. Role of the 2-amino group of deoxyguanosine in sequence recognition by EcoRI restriction and modification enzymes.
    J Biol Chem. 1977 Oct 25;252(20):7273-8 PMID: 332689
  14. Photo-cross-linking of protein-nucleic acid complexes.
    Methods Enzymol. 1977;46:168-80 PMID: 333226
  15. Direct and specific photochemical cross-linking of adenosine 5'-triphosphate to an aminoacyl-tRNA synthetase.
    Biochemistry. 1977 Oct 18;16(21):4678-84 PMID: 334247
  16. Exposure of tryptophanyl residues and protein dynamics.
    Biochemistry. 1977 Dec 13;16(25):5546-51 PMID: 921949
  17. Replication of bacteriophage M13. XII. In vivo cross-linking of a phage-specific DNA binding protein to the single-stranded DNA of bacteriophage M13 by ultraviolet irradiation.
    J Mol Biol. 1977 Sep;115(1):45-59 PMID: 336896
  18. Photochemical cross-linking studies on the interaction of Escherichia coli RNA polymerase with T7 DNA.
    Biochemistry. 1978 Jul 25;17(15):2954-61 PMID: 359036
  19. DNA strand specificity in promoter recognition by RNA polymerase.
    Nucleic Acids Res. 1980 Dec 11;8(23):5895-912 PMID: 7008032
  20. Variables affecting the selectivity and efficiency of retention of DNA fragments by E. coli RNA polymerase in the nitrocellulose-filter-binding assay.
    Gene. 1981 Jan-Feb;13(1):75-87 PMID: 7016668
  21. Protein measurement with the Folin phenol reagent.
    J Biol Chem. 1951 Nov;193(1):265-75 PMID: 14907713
  22. Sedimentation rate as a measure of molecular weight of DNA.
    Biophys J. 1963 Jul;3:309-21 PMID: 14016998
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/10.7.2399
Published
1982-04-10
Pages
2399-414
Language
English
Region
England
NLM ID
0411011
PMCID
PMC320618
Subset
IM
Grants
NIGMS NIH HHS · GM 00221 · United States
NIGMS NIH HHS · GM 21782 · United States
NIGMS NIH HHS · GM 28533 · United States
External Links
PubMed source DOI: 10.1093/nar/10.7.2399 PMC Full Text View journal details
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